Repeated Dose 4-Week Oral-Treatment for DRF Toxicity Test of HMC05 in Sprague-Dawley Rats

서 론 . HMC05 (B030007, B050042)

,

, , , , , , , ,

Repeated Dose 4-Week Oral-Treatment for DRF Toxicity Test of HMC05 in Sprague-Dawley Rats

Heung-Mook Shin

Department of Physiology, College of Oriental Medicine, Dongguk University

Objectives: HMCO5 is an extract obtained from 8 different herbal mixtures. We undertook a safety evaluation of HMCO5 for a dose range finding (DRF) toxicity test in specific pathogen free (SPF) Sprague-Dawley (SD) male and female rats.

Methods: The male and female rats were divided into 4 groups, respectively; G(0), treated with distilled water: G(1), treated with 222 mg/kg HMC05: G(2), treated with 667 mg/kg HMC05, and G(3), treated with 2,000 mg/kg HMC05;

HMC05 was administered orally for 4 weeks. The safety evaluation examined clinical signs, mortality, body weight, food consumption, water consumption, ophthalmic findings, urinalysis, hematological values, absolute & relative organ weights, and necropsy findings during the tests.

Results: There were no changes in clinical signs, mortality, body weight, food consumption, water consumption, and ophthalmic findings examined during the test periods. In serum biochemical values, triglyceride was increased in male group G(3) and Na

decreased significantly in male groups G(2), G(3) and G(4). In male group G(4), spleen weight decreased relatively and increases of absolute & relative left ovary weights were found. In addition, an adhesion of liver to diaphragm was found in male group G(2). However, we could not find any dose-interrelationships in these changes.

Conclusions: These results indicate that HMC05 extract did not show any toxicity in the DRF toxicity study.

Therefore, it suggests that establishment of 1,000, 333 and 111 mg/kg dosages are moderate in a repeated dose 26-week oral toxicity study of HMC05.

Key Words : HMC05, DRF toxicity, safety evaluation, dose-interrelationships 신흥묵

동국대학교 한의과대학 생리학교실

Original Article

Received：10 June 2009 Revised：22 July 2009 Accepted：24 July 2009

Correspondence to：신흥묵(Heung-Mook Shin) 경북 경주시 석장동 707 동국대학교 한의과대학

Tel：+82-54-770-2372, Fax：+82-54-742-5441, E-mail：[email protected]

1,2)

. HMC05

3)

, apoE

4)

, HPLC/UV LC-MS hesperidin, coptisine, palmatine berberine

.

2,000mg/kg (ALD)

6)

.

2005-60 (2005

10 21 ) ‘ ’

2005-79 (2005 12 21 )

‘ ’

HMC05 Sprague-

Dawley 4

.

HMC05

2,000 /

3 HMC05 222, 667

2.000 / /day 1 1 4

, ,

, , , ,

,

.

실험재료 및 방법

1. 시험물질

HMC05 ( 1.5 : 2

: 1 : 1 : 1.5 : 1.5 : 1.5 : 1.5 ; Table 1) ( ) Bioland ( , )

. 100 1 30

(1µ ) ( 60-70 cmHg)

(100’C) 1 1

1 .

51.5% .

2. 시험동물

HsdKoat:Sprague

Dawley

SD

SPF( )

5 24 .

,

,

.

( , )

,

. 7 ,

. .

3. 사육환경

1)

2

Pharmaceutical name Scientific name Ratio

半夏 Pinelliae ternate Ten. Ex Breitenb. 1.5

白朮 Atractylodes macrocephala Koidz. 2

天麻 Gastrodia elata Blume 1

陳皮 Citrus unshiu Marcow. 1

茯 Poria cocos Wolf 1.5

山 Crataegus pinnatifida Bunge var. typica C.K. Schneider 1.5

稀 Siegesbeckia pubescens Makio. 1.5

黃連 Coptidis japonica Makino 1.5

Table 1.

6 . 23±

3 , 55±15%, 10 20 /hr,

12 ( 8 8 )

150 300Lux .

(121 , 20 ) . 2)

(TEKLAD CERTIFIED GLOBAL 18% PROTEIN RODENT DIET, 2918C, Harlan Co. Ltd., USA)

.

, .

SOP .

3) ,

(W 215 × L

355 × H 200mm) 5

/ , 3 /

. 4)

ear-punch .

, .

, , ,

, .

4. 시험군 및 투여방법

1)

G1 ,

6)

2,000 /

, ,

2,000 /

3 HMC05

222, 667 2.000 / /day G2, G3,

G4 . 5

10 .

군 성 별 동물수

(마리) 동 물 번 호 투여액량

(ml/kg/day)

투여량 (mg/kg/day) G1

M / F 5 / 5 1-5 / 21-25 10 0

G2 M / F 5 / 5 6-10 / 26-30 10 222 G3 M / F 5 / 5 11-15 / 31-35 10 667 G4 M / F 5 / 5 16-20 / 36-40 10 2,000

2)

,

. 10 / ,

7 / , 4 .

5. 관찰 및 검사 항목

1)

,

, 1 1

. 2)

,

1 .

. 3)

1 ,

,

,

(g/rat/day) .

4)

.

5

, ( , ,

Lot 016360) ,

(Genesis, Gowa Co. Ltd., Japan)

, . ,

. 5)

(glucose), (bilirubin), (ketone body), (specific gravity), (occult

blood), (pH), (protein), (uro-

bilinogen), (nitrite) (leukocyte) (Multistix 10SG, Bayer) (CliniTek 100, Bayer)

. 6)

ether

.

/ , , ,

, . 7)

1

EDTA-2K CBC bottle

(ADVIA 2120, SIEMENS, USA) .

8)

clot activator 5ml vacutainer tube (IMPROVE, IMPROVE,

Germany) 15 20

10 (Hanil MF300, Korea,

3,000 rpm, 1,630 RCF) AU400

(Olympus, Japan) ,

(644 Na+/K+/Cl-Analyzer, SIEMENS, USA and RAPIDCHEM 744 Na+/K+/Cl-Analyzer,SIEMENS,

USA) .

항 목 단 위 방 법

ⓐ AST (Aspartate aminotransferase) U/L IFCC법

ⓑ ALT (Alanine aminotransferase) U/L IFCC법

ⓒ ALP (Alkaline phosphatase) U/L P-NPP법

ⓓ BUN (Blood urea nitrogen) mg/dL Urease-UV법

ⓔ CRE (Creatinine) mg/dL Jaffe법

ⓕ GLU (Glucose) mg/dL UV법

ⓖ CHO (Total cholesterol) mg/dL Enzyme법

ⓗ PRO (Total protein) g/dL Biuret법

ⓘ CPK (Creatine phosphokinase) U/L IFCC법

ⓙ ALB (Albumin) g/dL BCG법

ⓚ BIL(Total bilirubin) mg/dL Evelyn-Malloy법

ⓛ TG (Triglyceride) mg/dL Enzyme법

ⓜ IP (Inorganic phosphorus) mg/dL UV법

ⓝ Ca2+(Calciumion) mg/dL O-CPC 법

ⓞ A/G ratio (Albumin/Globulin

ratio) ratio PRO, ALB로 산출

ⓟ Na+(Sodiumion) mmol/L 전극법

ⓠ K+(Potassiumion) mmol/L 전극법

ⓡ Cl-(Chlorideion) mmol/L 전극법

ⓐ-ⓞ: 혈액생화학분석기(AU400, Olympus, Japan)를 이용하여 측정.

ⓟ-ⓡ: 전해질자동분석기(644 Na+,K+,Cl-Analyzer,SIEMENS,USA)로 측정.

항 목 단 위 측 정 방 법

ⓐ WBC (White blood cell

count) 103/μL Flowcytometry

ⓑ RBC (Red blood cell count) 106/μL Flowcytometry, Isovolumetry

ⓒ HGB (Hemoglobin conc.) g/dL Modified CN met-Hb method

ⓓ HCT (Hematocrit) % (RBC×MCV)÷10

ⓔ MCV (Mean corpuscular

volume) fL Histogram

ⓕ MCH (Mean corpuscular

hemoglobin) pg (HGB÷RBC)×10

ⓖ MCHC (Mean corpuscular

Hb conc.) g/dL [HGB÷(RBC×MCV

)]×1000

ⓗ RDW (Red cell distribution

width) % Histogram

ⓘ HDW (Hb conc. distributio

n width) g/dL Histogram

9)

, , , , , ,

, , , , ,

(BP221S and CP153, Sartorius AG, Germany) ,

.

.

6. 통계처리

(parametric multiple comparison procedures)

(non parametric multiple comparison proc-

edures) ,

SPSS 10.1

. , ,

,

one-way ANOVA test

, Levene

test ,

T-test .

Kruskal-Wallis’ H-test .

척도 변환

GLU, KET, PRO,BIL,

OB, WBC SG pH URO NIT

0 - ≤1.005 6.5 0.1 -

1 +/- 1.010 7.0 1.0 +

2 1+ 1.015 7.5

3 2+ 1.020 8.0

4 3+ 1.025 8.5

5 4+ ≥9.0 ≥9.0

결 과

1)

,

(Table 2).

2) HMC05

(Table 3).

3)

HMC05

(Table 4).

4)

HMC05

(Table 5).

5)

HMC05 (Table 6).

6)

HMC05

(Table 7).

7)

HMC05

(Table 8).

Groups (mg/kg/day) Sex: male or female

Days Observed G1 (0) G2 (222) G3 (667) G4 (2,000)

0 ~ 28 Normal 5 / 5

5 / 5 5 / 5 5 / 5

28 Terminal sacrifice 5 / 5 5 / 5 5 / 5 5 / 5

*, Data are the number of animals with the sign/Number of animals examined.

Table 2.

BODY WEIGHT CHANGES (g) Group (mg/kg) / Sex (male/female)

Weeks G1 (0) G2 (222) G3 (667) G4 (2,000)

0 161.94±5.10 / 124.68±2.80 162.69±3.12 / 125.80±4.12 161.96±7.16 / 125.93±4.78 163.32±4.50 / 126.76±5.03 1 220.29±8.95 / 150.93±4.52 219.49±5.73 / 153.57±6.65 223.75±7.95/ 153.08±11.81 222.73±6.06 / 149.68±10.39 2 268.01±14.56 / 171.99±5.77 268.43±4.44 / 153.57±6.65 276.74±12.27 / 171.48±14.02 277.04±9.41 / 172.92±10.16 3 306.55±16.34 / 191.90±5.30 308.69±4.19 / 175.11±9.15 316.52±17.31 / 187.04±15.63 317.36±14.43 / 187.23±11.76 4 332.50±18.49 / 202.27±5.05 333.90±4.20 / 201.15±6.56 345.14±21.47 / 201.35±20.53 339.28±16.78 / 201.06±12.12 Weight Gains 170.55±16.64 / 77.59±5.55 171.21±6.06 / 75.36±2.69 183.17±19.00 / 201.35±20.53 175.96±16.30 / 201.06±12.12

N 5 / 5 5 / 5 5 / 5 5 / 5

Table 3.

FOOD CONSUMPTIONS (g) Group (mg/kg) / Sex (male/female)

Weeks G1 (0) G2 (222) G3 (667) G4 (2,000)

0 21.80±0.68 / 16.80±0.29 22.18±1.17 / 17.34±0.16 22.78±1.09 / 16.74±1.42 23.70±0.54 / 6.21±0.69 1 23.27±2.06 / 14.43±1.11 24.92±0.76 / 16.32±2.12 25.44±2.53 / 14.41±3.59 26.05±0.81 / 16.31±0.81 2 25.46±0.13 / 16.60±1.14 25.60±1.00 / 17.34±0.43 25.51±1.75 / 17.13±1.28 24.62±2.44 / 16.67±1.31 3 22.94±0.28 / 12.42±1.05 23.75±0.54 / 14.02±1.56 24.05±3.94 / 13.09±2.72 23.74±0.13 / 14.97±0.88 4 27.66±0.52 / 18.05±1.25 27.83±2.09 / 18.02±2.52 25.57±3.28 / 17.93±4.89 26.00±0.29 / 17.18±0.21

N 5 5 5 5

Table 4.

WATER CONSUMPTIONS (g) Group (mg/kg) /Sex (male/female)

Weeks G1 (0) G2 (222) G3 (667) G4 (2,000)

0 21.94±1.55 / 19.41±0.38 21.84±1.64 / 20.22±0.88 23.62±0.72 / 18.14±0.03 25.94±1.01 / 18.12±0.57 1 25.53±4.78 / 16.91±3.15 26.00±0.91 / 20.10±4.09 28.79±0.83 / 15.65±4.29 30.05±6.21 / 18.64±1.49 2 30.08±2.81 / 23.48±2.43 28.98±3.24 / 24.85±1.83 32.69±0.23 / 20.97±3.04 29.27±5.67 / 22.69±0.81 3 26.15±1.83 / 18.12±0.70 24.68±0.87 / 17.96±1.14 28.96±2.18 / 15.72±4.16 28.44±6.39 / 19.22±0.56 4 30.06±1.12 / 28.30±3.07 28.14±1.25 / 25.49±2.06 29.52±0.75 / 24.36±4.15 29.00±6.46 / 24.37±4.35

N 5 / 5 5 / 5 5 / 5 5 / 5

Table 5.

OPTHALMIC FINDINGS Group (mg/kg/) / Sex (male or female)

Sites Findings G1 (0) G2 (222) G3 (667) G4 (2,000)

Left eye No gross findings 5 / 5* 5 / 5 5 / 5 5 / 5

Right eye No gross findings 5 / 5 5 / 5 5 / 5 5 / 5

*, Number of animals with the sign / Number of animals examined

Table 6.

URINALYSIS

Groups (mg/kg/day)

Test

Items Result Grade

Male Female

G1 G2 G3 G4 G1 G2 G3 G4

0 222 667 2,000 0 222 667 2,000

GLU - 0 5 5 5 5 5 5 5 5

BIL - 0 5 5 5 5 5 5 5 5

KET 　

- 0 3 3 2 0 5 5 4 2

+/- 1 2 2 3 4 0 0 1 3

1+ 2 0 0 0 1 0 0 0 0

SG 　

≤1.005 0 0 0 0 0 0 0 0 1

1.010 1 0 0 0 0 1 0 0 0

1.015 2 0 0 0 0 2 5 1 0

1.020 3 1 0 0 0 0 0 2 0

1.025 4 1 3 0 0 2 0 1 3

1.030 5 3 2 5 5 0 0 1 1

pH 　

7.0 0 2 0 0 0 0 0 0 0

7.5 1 3 5 5 5 3 1 2 1

8.0 2 0 0 0 0 2 4 3 3

8.5 3 0 0 0 0 0 0 0 1

PRO　

+/- 0 0 0 0 0 0 1 1 1

1+ 1 0 0 0 0 2 2 1 1

2+ 2 1 0 1 0 2 2 1 1

3+ 3 4 5 4 5 1 0 2 2

URO 0.1 0 5 5 5 5 5 5 5 5

NIT - 0 5 5 5 5 5 5 5 5

OB

- 0 3 3 4 3 5 5 5 4

+/- 1 1 0 0 1 0 0 0 0

1+ 2 1 2 0 1 0 0 0 0

2+ 3 0 0 0 0 0 0 0 0

3+ 4 0 0 1 0 0 0 0 1

WBC

- 0 0 1 0 0 1 0 1 3

+/- 1 2 1 0 2 2 2 2 2

1+ 2 2 3 2 3 2 3 2 0

2+ 3 1 0 3 0 0 0 0 0

N 5 5 5 5 5 5 5 5

GLU: Glucose, BIL: Bilirubin, KET: Ketone body, SG: Specific gravity, PRO: Protein, NIT: Nitrite, URO: Urobilinogen, OB: Occult blood, WBC: Leukocyte

Table 7.

SERUM BIOCHEMICAL VALUES Groups (mg/kg/day) / Sex (male/female)　

Tests Units G1 (0) G2 (222) G3 (667) G4 (2,000)

AST U/L 101.9±32.3 / 90.1±8.7 88.6±9.5 / 79.6±6.1 96.0±11.6 / 83.1±2.6 84.9±1.8 / 92.8±12.0 ALT U/L 53.0±19.5 / 35.9±2.4 50.9±13.7 / 34.7±3.8 41.8±4.9 / 35.1±4.4 36.3±2.7 / 34.9±4.9 ALP U/L 152.8±36.5 / 110.9±7.1 145.6±18.0 / 106.8±13.8 148.1±27.4 / 104.6±18.7 152.5±29.3 / 91.8±9.0 BUN mg/dL 18.9±3.0 / 20.1±3.0 17.9±1.5 / 18.9±1.3 17.1±1.6 / 21.9±2.7 16.1±1.2 / 18.9±2.0 CRE mg/dL 0.48±0.09 / 0.53±0.05 0.52±0.03 / 0.53±0.05 0.51±0.03 / 0.55±0.04 0.52±0.03 / 0.52±0.02 GLU mg/dL 127.6±10.2 / 105.7±11.0 127.7±11.0 / 109.2±15.5 120.4±12.8 / 106.7±13.6 128.8±11.5 / 102.6±3.5 CHO mg/dL 111.2±22.9 / 109.2±15.6 111.2±9.4 / 96.0±11.4 101.6±21.2 / 104.4±21.7 107.0±14.1 / 95.8±25.8 PRO g/dL 6.13±0.26 / 6.05±0.18 6.23±0.22 / 6.09±0.24 6.24±0.20 / 6.17±0.33 6.26±0.13 / 6.10±0.22 CPK U/L 159.2±122.3 / 157.0±78.1 133.0±46.7 / 92.4±30.7 384.0±406.7 99.8±6.3 133.6±34.0 / 125.0±59.3 ALB g/dL 3.04±0.32 / 3.20±0.07 3.09±0.26 / 3.23±0.13 3.12±0.11 / 3.23±0.15 3.21±0.12 / 3.31±0.09

BIL mg/dL 0.15±0.02 / 0.18±0.01 0.16±0.01 / 0.18±0.02 0.16±0.01 / 0.18±0.01 0.16±0.01 / 0.17±0.01 TG mg/dL 52.0±3.3 / 41.4±3.5 45.4±5.6 / 42.2±5.1 51.0±7.5 / 47.2±14.2 64.4±8.5

/ 44.0±10.1 IP mg/dL 8.06±0.59 / 7.36±0.66 8.06±0.46 / 7.69±0.33 8.14±0.55 / 7.08±0.32 8.26±0.45 / 7.52±0.35 Ca2+ mg/dL 10.35±0.21 / 10.04±0.19 10.04±0.19 / 10.08±0.18 10.10±0.15 / 10.04±0.17 10.32±0.21 10.05±0.33

A/G ratio 0.99±0.13 / 1.13±0.08 0.99±0.14 / 1.13±0.09 1.00±0.05 / 1.11±0.08 1.05±0.06 / 1.19±0.05 Na+ mmol/L 142.2±0.4 / 140.4±0.4 141.4±0.7

/ 140.3±0.8 141.5±0.7

/ 140.4±1.4 141.1±0.2

/ 140.2±0.6 K+ mmol/L 4.25±0.12 / 4.25±0.18 4.32±0.16 / 4.24±0.46 4.42±0.19 / 3.91±0.17 4.51±0.09 / 4.24±0.35 Cl- mmol/L 103.7±1.2 / 104.9±0.6 102.8±0.3 / 104.7±1.1 102.8±1.5 / 104.2±0.7 103.1±0.5 / 104.3±0.5

N 5 5 5 5

*/** Represents a significant difference at p<0.05/p<0.01 level compared with the vehicle control.

AST: Aspartate aminotransferase, ALT: Alanine aminotransferase, ALP: Alkaline phosphatase, BUN: Blood urea nitrogen, CRE: Creatinine, GLU: Glucose, CHO: Total cholesterol, PRO: Total protein, CPK: Creatine phosphokinase, ALB: Albumin, BIL: Total bilirubin, TG: Triglyceride, IP: Inorganic phosphorus, Ca2+: Calciumion, A/G ratio: Albumin/Globulin ratio, Na+: Sodiumion, K+: Potassiumion, Cl-: Chlorideion.

Table 9.

HEMATOLOGICAL VALUES Groups (mg/kg/day) /Sex (male/female)

Tests Units G1 (0) G2 (222) G3 (667) G4 (2,000)

WBC 103/ μL 10.57±1.43 / 4.81±1.18 10.82±1.0 / 6.04±1.74 8.97±0.60 / 5.27±1.86 11.55±2.31 / 4.63±0.44 RBC 106/ μL 8.14±0.39 / 7.46±0.36 8.07±0.17 / 7.55±0.25 7.78±0.33 / 7.72±0.23 7.80±0.23 / 7.51±0.22 HGB g/dL 15.4±0.5 / 14.5±0.5 15.3±0.5 / 14.6±0.3 15.0±0.3 / 14.7±0.4 15.1±0.4 / 14.9±0.3 HCT % 47.0±1.6 / 42.6±1.5 46.6±1.3 / 42.8±1.5 46.0±1.3 / 43.3±0.9 45.7±1.3 / 43.6±1.5 MCV fL 57.9±2.2 / 57.2±1.3 57.8±0.7 / 56.6±1.3 59.2±0.9 / 56.1±1.9 58.6±2.0 / 58.0±0.7 MCH pg 18.9±0.6 / 19.5±0.7 19.0±0.2 / 19.3±0.4 19.4±0.5 / 19.0±0.9 19.4±0.6 / 19.9±0.2 MCHC g/dL 32.8±0.4 / 34.1±0.4 32.9±0.2 / 34.1±0.7 32.7±0.3 / 33.9±0.4 33.1±0.2 / 34.3±0.7 RDW % 10.8±0.3 / 10.4±0.3 10.8±0.1 / 10.2±0.2 11.1±0.3 / 10.3±0.2 11.1±0.4 / 10.4±0.2 HDW g/dL 2.46±0.15 / 2.36±0.12 2.48±0.18 / 2.33±0.12 2.43±0.15 / 2.39±0.19 2.37±0.06 / 2.28±0.10

N 5 / 5 5 / 5 4 / 5 5 / 4

WBC: White blood cell count, RBC: Red blood cell count, HGB: Hemoglobin conc., HCT: Hematocrit, MCV: Mean corpuscular volume, MCH: Mean corpuscular hemoglobin, MCHC: Mean corpuscular Hb conc., RDW: Red cell distribution width, HDW: Hb conc. distribution width.

Table 8.

8)

2,000 / TG

(p<0.01)

, 222, 667 2,000 / Na+

(p<0.05)

.

(Table 9).

9)

4 HMC05

ABSOLUTE & RELATIVE ORGAN WEIGHTS (g)

Groups (mg/kg/day) G1 (0) G2 (222) G3 (667) G4 (2,000)

Body weight

304.01±19.55 304.15±4.82 317.83±16.73 312.96±13.78

Adrenall Gland-Left 0.0244±0.0007 0.0226±0.0027 0.0246±0.0013 0.0248±0.0031

% to Body weight 0.0080±0.0004 0.0074±0.0009 0.0078±0.0004 0.0079±0.0010 Adrenall Gland-right 0.0241±0.0031 0.0213±0.0020 0.0210±0.0052 0.0239±0.0035

% to Body weight 0.0079±0.0010 0.0070±0.0006 0.0067±0.0019 0.0076±0.0011

Thymus 0.4824±0.0449 0.4712±0.0397 0.5346±0.0609 0.5090±0.1003

% to Body weight 0.1592±0.0180 0.1549±0.0122 0.1680±0.0141 0.1621±0.0277

Prostate 0.3138±0.0881 0.3176±0.0659 0.3872±0.0414 0.3736±0.0677

% to Body weight 0.1035±0.0288 0.1043±0.0208 0.1218±0.0107 0.1192±0.0193

Tests-Left 1.7802±0.0936 1.8664±0.0835 1.8764±0.0494 1.8646±0.0827

% to Body weight 0.5860±0.0130 0.6140±0.0344 0.5916±0.0320 0.5968±0.0378

Testis-Right 1.8086±0.1084 1.8436±0.1246 1.8934±0.0688 1.8620±0.0811

% to Body weight 0.5957±0.0315 0.6064±0.0440 0.5965±0.0237 0.5957±0.0322

Epididymis-Left 0.4064±0.0200 0.4044±0.0387 0.4128±0.0289 0.4522±0.0263

% to Body weight 0.1338±0.0047 0.1329±0.0123 0.1299±0.0074 0.1449±0.0130

Epididymis-Right 0.4142±0.0309 0.4200±0.0447 0.4218±0.0218 0.4342±0.0135

% to Body weight 0.1363±0.0083 0.1381±0.0145 0.1328±0.0044 0.1389±0.0064

Spleen 0.7720±0.1122 0.7034±0.0503 0.7364±0.0731 0.6744±0.0420

% to Body weight 0.2533±0.0258 0.2312±0.0144 0.2316±0.0179 0.2154±0.0079

Kidney-Left 1.0446±0.0440 0.9868±0.0425 1.1200±0.0970 1.0740±0.0729

% to Body weight 0.3449±0.0287 0.3245±0.0146 0.3520±0.0146 0.3433±0.0208

Kidney-Right 1.0818±0.0714 1.0282±0.0364 1.1180±0.0620 1.1084±0.0924

% to Body weight 0.3567±0.0289 0.3382±0.0149 0.3520±0.0145 0.3541±0.0235

Heart 1.1042±0.0638 1.0762±0.0592 1.1900±0.0770 1.1658±0.1099

% to Body weight 0.3637±0.0195 0.3539±0.0198 0.3742±0.0066 0.3722±0.0262

Lung 1.4572±0.1014 1.3414±0.0823 1.4166±0.0979 1.5568±0.1566

% to Body weight 0.4804±0.0391 0.4411±0.0267 0.4456±0.0167 0.4971±0.0398

Liver 9.7054±0.8296 9.3272±0.5910 9.9068±0.9460 9.2778±0.5194

% to Body weight 3.1918±0.1736 3.0649±0.1503 3.1155±0.2307 2.9653±0.1256

N 5 5 5 5

*, Represents body weights right before necropsy after fasting.

, Represents a significant difference at p<0.01 level compared with the vehicle control.

Table 10-1.

. HMC05 (222, 667, 2.000mg/kg/day)

. 2,000 mg/kg

(p<0.01) , 2,000mg/kg

(p<0.05) (Table 10-1, 10-2).

10)

(rubefaction) 1 , 667mg/kg

(adhesion of diaphragm) 1 . (clear liquid

content) 1 ,

(Table 11-1, 11-2).

ABSOLUTE & RELATIVE ORGAN WEIGHTS (g)

Groups (mg/kg/day) G1 (0) G2 (222) G3 (667) G4 (2,000)

Body Weight

189.39±4.12 186.82±6.57 186.53±19.03 185.41±10.97

Ovary-Left 0.0393±0.0029 0.0351±0.0058 0.0385±0.0049 0.0461±0.0052

% to Body weight 0.0208±0.0016 0.0189±0.0034 0.0208±0.0033 0.0248±0.0015

Ovary-Right 0.0413±0.0094 0.0387±0.0033 0.0394±0.0042 0.0445±0.0058

% to Body weight 0.0218±0.0049 0.0207±0.0021 0.0212±0.0023 0.0240±0.0018 Adrenel Gland-Left 0.0305±0.0037 0.0295±0.0029 0.0313±0.0029 0.0307±0.0019

% to Body weight 0.0161±0.0017 0.0158±0.0017 0.0169±0.0019 0.0166±0.0013 Adrenel Gland-Right 0.0291±0.0042 0.0290±0.0021 0.0300±0.0024 0.0299±0.0035

% to Body weight 0.0153±0.0020 0.0156±0.0014 0.0162±0.0014 0.0162±0.0021

Thymus 0.3312±0.0481 0.3406±0.0443 0.3540±0.0722 0.3562±0.0228

% to Body weight 0.1746±0.0229 0.1825±0.0248 0.1885±0.0221 0.1924±0.0118

Uterus 0.4184±0.0537 0.6294±0.5246 0.4868±0.2591 0.3982±0.1195

% to Body weight 0.2207±0.0260 0.3375±0.2812 0.2610±0.1319 0.2129±0.0523

Spleen 0.5164±0.0181 0.5174±0.0682 0.5536±0.0816 0.4948±0.0684

% to Body weight 0.2727±0.0096 0.2767±0.0338 0.2970±0.0344 0.2664±0.0266

Kidney-Left 0.6512±0.0249 0.6236±0.0189 0.6390±0.0488 0.6598±0.0342

% to Body weight 0.3439±0.0144 0.3341±0.0156 0.3435±0.0171 0.3560±0.0071

Kidney-Right 0.6640±0.0324 0.6450±0.0292 0.6666±0.0516 0.6748±0.0572

% to Body weight 0.3508±0.0210 0.3456±0.0192 0.3582±0.0140 0.3641±0.0251

Heart 0.8148±0.0133 0.7978±0.0488 0.8004±0.0953 0.7978±0.0890

% to Body weight 0.4303±0.0058 0.4270±0.0193 0.4301±0.0418 0.4293±0.0234

Lung 1.0822±0.0345 1.0606±0.0588 1.0722±0.0811 1.1032±0.0635

% to Body weight 0.5717±0.0229 0.5681±0.0338 0.5780±0.0530 0.5957±0.0315

Liver 5.5218±0.2731 5.3120±0.1942 5.3486±0.8205 5.3742±0.4621

% to Body weight 2.9155±0.1273 2.8440±0.0757 2.8618±0.2707 2.8950±0.0850

N 5 5 5 5

*, Represents body weights right before necropsy, after fasting

, Represents a significant difference at p<0.05 level compared with the vehicle control

Table 10-2.

고 찰

HMC05

( 10-0577674 / 10-0787174 )

HMC05 . HMC05

3)

, apoE

4)

hesperidin, coptisine, palmatine

berberine

.

. 2003-17

HMC05

6)

4

26

. HMC05 222, 667

2,000mg/kg SD

10 ( 5 ) 1 1 , 4

, , , ,

, , ,

.

4

, .

, ,

. 2,000mg/kg

TG 222, 667, 2.000mg/kg

Na

.

11,12)

. 2,000

mg/kg .

, ,

. , 2,000mg/kg

, HMC05 . Groups (mg/kg/day)

Organs Signs G1 (0) G2 (222) G3 (667) G4 (2,000)

Thymus Rubefaction 1 (20)

- - -

Liver Adhesion of diaphragm - - 1 (20) -

N 5 5 5 5

*, Number of animals with the sign (%). -, No necropsy finding was found.

Table 11-1.

Groups (mg/kg/day)

Organs Signs G1 (0) G2 (222) G3 (667) G4 (2,000)

Uterus Clear liquid content 1 (20)

- - -

N 5 5 5 5

*, Number of animals with the sign (%). -, No necropsy finding was found.

Table 11-2.

1 ,

13,14)

.

667mg/kg 1

.

, HMC05

. HMC05

,

TG Na

HMC05 4

SD .

결 론

HMC05 4 SD

DRF .

, , , ,

,

. HMC05 2,000mg/kg

TG Na

, .

2,000mg/kg ,

667mg/kg 1

.

HMC05 26

OECD guideline 1,000mg/kg

, 3

333mg/kg , 111mg/kg

.

감사의 글

(B080031)

.

참고문헌

1. Lee HJ, Seong YJ, Kim SD, Moon KJ, Kim JB, Kim GW, et al. Enhanced vasorelaxation of BanhabackchulChunma-Tang and involved mechanism. Korean J. Oriental Physiology &

Pathology. 2005;19(5):1311-1316.

2. Korean intellectual property office: A curing and preventing pharmaceutical composition having an effect of anti-hypertension inducing by cont- aining an extracts of chinese herb, Registration No. 10-0577674 (May 1, 2006)

3. Moon KJ, Jang HO, Kim GW, Shin HM. Signaling mechanism on the vascular relaxation of HMC05.

Korean J. Oriental Physiology & Pathology.

2008;22(2):315-320.

4. Kim KM, Choi JY, Yoo SE, Park MY, Lee BS, Ko YH, et al. HMCO5, Herbal Extract, Inhibits NF-κB Expression in Lipopolysaccharide Treated Macrophages and Reduces Atherosclerotic Lesions in Cholesterol Fed Mice. Journal of Ethnopha- rmacology. 2007;114:316-324.

5. Kim SH, Choi EJ, Lee KY, Sung SH, Shin HM.

Simultaneous determination of alkaloids and flavonoids in HMC05 preparation by HPLC- DAD. Journal of Liquid Chromatography &

Related Technologies, Ethnopharmacology. 2008

;31:2917-2926.

6. Shin HM. Acute Oral Toxicity of HMC05 in SD Rats. Korean J. Oriental Physiology & Pathology.

2008;22(6):1562-1565.

7. Korea Food and Drug Administration: Toxicity

Test Guidelines for Safety Evaluation of Drugs,

Notification No. 2005-60 (Oct. 21, 2005).

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9. OECD. OECD Guidelines for the Testing of chemicals. TG 401 & 420. 2002.

10. Hayes AW. Principles and Methods of Toxicology.

2

Edition. New york:Raven Press. 2001:177.

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Animal Science. 1990;6(1):33-43

12. Kang BH, Son HY, Ha CS, Lee HS, Song SW.

Reference Values of Hematology and Serum Chemistry in Ktc:Sprague-Dawely Rats. The Korean Journal of Laboratory Animal Science.

1995;11(2):141-145

13. Greaves P. Histopathology of preclinical toxicity studies; Interpretation and relevance in drug evaluation. Elsevier. 2000

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CA, Jr WF, Mackenzie. Pathology of the Fischer

Rat. Academic Press. 1990.